When search the mechanism of blood clotting, important 8 factors interfering biochemical reaction can be considered. To speed up the formation of fibrin is important to coagulate of incision and open wound. Therefore two effective process can be examined: inhibition of plasmin enzyme and providing the calcium ion flow. Calcium salts or fibrin formation with anti-fibrinolytic compound and stabilisation of fibrin can be obtained (S. Samudrala, Aorn J. Inc. September V.88 (3) 2008).
The basic aim of this study are to produce the bandage, pad, powder and gel with haemostatic and antiseptic features and release the medical market.
Dry cellulose based hemostatic bands or pads is impregnated by some substances formed solid, powder or gel, featured hemostatic and anti-septic, will be designed to control bleeding on traumatic and severe bleeding.
To prevent the bleeding of open wound and incision, it is important to provide the formation of fibrin and the stabilisation of fibrin. The coagulation process and its steps are shown on Schema-1. Two important steps are inhibition of fibrinolysis enzymes and sufficient calcium ions flow is activated Factor XII. Therefore fibrin formation with Ca+2 ion and with anti-fibrinolytic (anti-plasmin) as like Aprotinine, 6-aminocaproic acid, 6-acetaminocaproic acid, tranexamic acid, 4-aminomethyl benzoic acid and its stabilisation will be obtained (S. Samudrala, Aorn J. Inc. September V88 (3) 2008).
This study also comprise to design the anti-microbial protective products together with its benefits as controlling of bleeding for hemostatic purposes.

There are two steps for the hemostasis under the bleeding wounding, trauma.
Primer hemostasis means that vascular wall established any reason is formation of blood platelet in the result of thrombocyte local activation and aggregation. After wounding, thrombocytes bound collagen protein fibres; glycoprotein-Ib and plasma factors is necessary for this reaction. The platelets aggregate with homogen mass unless the wound is big. Under the external effect neuromediator as like ceratonine secrete and it causes the vasoconstriction and have the significant role for speeding up the coagulation. Shorter hemostatic time is the important point for this study.
Secondary hemostasis: Platelets formed at primer step is not enough for complete healing. To obtain sufficient hardness, fibrin formation in the other words coagulation thrombosis is required. For this formation there are intrinsic and extrinsic factors together with trombone generation on the platelet to change insoluble fibrin from soluble fibrin. The fibrin stabilisation factor is calcium ions at this step (Schema 1).
Changing soluble fibrin mechanism is essential to block fibrinolysis enzyme. It is the subject to add local effects along with anti-fibrinolytic compounds.
According to Morawitzs, for the coagulation there are 4 basic factors. “Farmasötik Kimya, Hacettepe Üniv. Eczacilik Fak. Yayinlar (2013), E. Mutschler, et. al. Arzneimittel wirkungen, Wiessenschaftliche Verlag Gmbh Stutgart 9. Auflage (2008)”                Factor I Fibrinogen        Factor II Prothrombin        Factor III Thromboplastin        Factor IV Calcium ion        
It is well known that the result of thrombin reaction with fibrinogen is obtained the clot; the formation of fibrin and joining of 8 various factors ensure to stop the bleeding. All coagulation factors are the proenzyme excluding Factor III. To stop bleeding there is two important points; first cellular which is blood platelet and second humoral factor which is formation of the fibrin. During trauma, on external and internal bleeding the formation of fibrin as humoral factor is mainly important.
The initial material and technology research shows there are not many alternatives for the preparation of pad, gauze, powder and gel products which have hemostatic, antiseptic properties. Indeed some commercial brands for example: Hemcon, Anscare, Ankaferd Blood Stopper (ABS), Surcicel, Dettol, have either hemostatic or antiseptic feature. This study aim is the methods of preparing more useful hemostatic, anti-septic compound and certificate with its in-vitro in-vivo tests (or new tests if requires). In other words, anti-fibrinolytic, hemostatic, hypoallergic (non cytotoxic), partial antimicrobial powder and pads for use in trauma and mid severe wound will be developed. Main aim is hemostatic time which will be in the range of 0.5-3.5 minutes.
To reach the aim mentioned above, formulation plan:    7. The formation of carrier matrix; the preparation of chitosan and oxidised cellulose based on hemostatic polymer    8. Impregnation of chemicals effective antimicrobial and anti-fibrinolytic on hemostatic polymer matrix material which is recorded on the literatures    9. Plaster this pad and packing against environmental effect    10. Sterilise by gamma radiation and compliance test for dermatologic, cytotoxic, microbiological at the university or accredited lab.    11. Determine the hemostatic time and antimicrobial minimal inhibition concentration (MIC) values in compliance with European Pharmacopeia in matrix material mentioned item 1.    12. Chemicals as antifibrinolytic effect plan to use is tranexamic acid and 6-aminocaproic acid and aprotinin.
The product after present invention, features must be:                Hemostatic time will be 0.5-3.5 min.        Large antimicrobial effective spectrum        Must be completed the in-vitro and in-vivo tests (hemostatic time of rat liver, cytotoxic effect, irritation, sensitisation, antimicrobial effect).        Must have hypo allergic feature        Soft and absorb the serum        Do not stick to the surface of the wound        Proof the compliance with biodegradable standards via histopathological animal tests.        Determine the bio absorption time must be 2-5 days        
Matrix Formation:
Until today hemostatic and antiseptic polymer matrix for use in medical purposes, had two basic targets:                Anionic matrix (oxidised cellulose and its derivates)        Cationic matrix (Chitosan, gelatine, fibril collagen and its derivates)        
1.1. Anionic Matrix Polymer:
As a result of various natural cellulose derivate chemically, obtain hydroxyethyl cellulose, 2-hydroxypropyl cellulose, methyl-ethyl-cellulose and carboxymethyl cellulose. This cellulose derivates obtain the substitution of OH groups on the glucose unit of the cellulose. These do not have any hemostatic feature and have water absorption, gel formation with micelle concentration. Oxidised cellulose is formed anionic hemostatic polymer in rate of carboxyl group on the 6. C which carry oxidise cellulose derivates.
These products are called as poly anhydroglycuronic acid (PAGA). Polymeric mass carries carbonyl groups in yield 8-30% of carboxyl. In general linear polymer has 23.6 or 30% in this yield depends on its branches of glucose polymer. The predicted rate of glycuronic acid in carboxyl molecule is more than 80%. During oxidation partly occur ketone carbonyl at 3 C. of aldehyde and glucose (Schema 2).

Aforesaid carbonyl have maximal 5% aldehyde carbonyl as request. More than this yield is not applicable for hemostatic polymer. In general desirable yield is less than 2.3%. After cellulose nitrate oxidation, anionic matrix contain maximal 0.5% bound nitrogen. In general desirable rate is less than 0.2%.
The size of carbohydrate polymer molecule size is 1×103-3×104 daltons. Regenerated cellulose molecule size is 1×103-5×103 that is bigger than linter cellulose. Viscon cellulose molecule size is the range of 5×103-1.5×104 dalton. Ideal size is 1500 k.dalton. Carbonyl amount of cellulose polymer is 12-26% of whole molecule. Glycuronic acid is 95% of it. In the molecule for the decreasing of aldehyde rate (if 0.6-1.5%) and increasing the yield of carboxyl is done by raw oxidised cellulose re-oxidised with hydrogenperoxite. Oxidised cellulose (which carry 16-22% carboxyl) and its salts as an anionic matrix are used for hemostatic purpose“ W. H. Ashton, U.S. Pat. No. 3,364,200 (1968)”.
1.2. Cationic Matrix Polymer:
Cationic matrix (polypeptide gelatine, fibrilar, collagen, 2-amino-D-glucose as polysaccharide and 2-acetamido-D-Glucose as chitosan) is used as hemostatic covers. They all are biocompatible hemostatic but some patients had occur allergic and erethitic reaction. Chitosan chitin is obtained by de-asetilation which 85% gain is necessary condition for hemostatic purpose.
For use in medical operation and external bleeding is subject to prepare intermolecular complex (IMC) from all these matrix. This invention aim is to use PAGA as a basic matrix, prefer to take either costing or toxicity side effects. Final product is dual effected medical material featured both flexible, absorbable, biocompatible hemostatic and antimicrobial.
Present invention try to determine the fabrication production method of powder, band, spanch, gel forms and IMC complex solution's preparation which specified above.
For powder formulation PAGA is quite sufficient. It is possible to prepare gel formulation with Gelatine and Chitosan. Biocompatible formulation is possible to add Ca and Na salts. Plaster band and spanch form production make difficulties to add these salts. Therefore latest studies start to use nonwoven textile or polymeric film. Animal hemostatic test and human tests are parallel relatively (0.99/0.01-0.01/0.99 statistically) for PAGA and this is its primer advantage. Secondly its hemostatic degree is measurable biologically. On the other hand its hemostatic time, bio-absorption time and immune modulative compliance values are measurable. The measurement techniques are written on pharmacopeias.
PAGA is compatible with polymeric cationic matrixes and use with. Cationic matrix groups are:    4. Biocompatible, with Nitrogen, Synthetic Oligomer and Polymer            a. Acrylamide and metacrylamide polymer and co-polymer, natural polysaccharides (gum-, quargum-, hydroxypropyl triammonium chloride etc.        b. Synthetic and semi-synthetic polyamino acid polilizin, polyarginine, -poli (N-(2-hydroxethyl) DL-asparamid and synthetic anti-fibrinolytic, hexadimethrine bromide (pyoben)            5. Natural and semi-synthetic peptides, gelatine, collogen, protamine, fibrinopeptide and its derivates.    6. Chitin as natural anti-aminoglucan and its fractions, its de-asetile derivatives are chitosan, microbial origin arthropods “crabs”
In this study these groups are not included because their measurement and standardisation have the difficulties and their cost is high.
As numerous literature and patent studies are recorded, PAGA matrix compounds/solutions was prepared with specified three groups above and could not reach satisfied results correspond to speed hemostatic and allergic and cytotoxic properties, stability at animal trials. IMC specified at 1a and 1b is tested preferably but only film and plaster had technic efficiency while working on non-woven, film, plaster and pad forms. Indeed prototype production (hemostatic and cover) is prepared for external wounding. We concentrate only oxidised cellulose production and its method standardisation on our invention. Second and third group cationic matrix are used in micro operations and need aseptic and automated systems to produce therefore this study focus on the low-cost production of biodegradable, biocompatible oxidised cellulose and its validation.
Regenerated oxidised cellulose (REOXC) is natural topic hemostatic biomaterial commonly used. It is formed that the selective oxidation of 6 C. primer alcohol groups on cellulose polymer. As a result of general oxidation, —COOH group is obtained and its range is 16-24% of total polymer. “Yadong Wu et al. Carbohydrate Polymers; 88 (2012) 1023-1032”. This product is comply to produce as sterilisable powder or textile manufacturing. This product is seen as powerful acute hemostatic and also it is known as great biosafety material in reference to “Zhu I. H. et. al. Journal of Pharmaceutics (2001), 223, 35-47”.
As it is known REOXC was produced by NO2 oxidation of cellulose on industrial level at 1930's (Lit. 5, 7, 13, 14). Selective oxidisation was done by “Yackel, E. C. and Kenyon, W.O. U.S. Pat. No. 2,232,990” at 1940's and analyse the oxidation kinetic and the rate of NO2/cellulose. The result of them is shown at Table-1. The more gas level in a unit volume is the more oxidation level of the cellulose.
TABLE 172 hours, NO2/Cellulose ratio and oxidation relation (Previous Method)NO2-Cotton concentrationfactor—COOH %0.6013.260.7513.380.9015.841.2019.801.5021.80
Many researcher have studied with different oxidants of this product and one of them is Proskulo et. al. carbohydrate Polymers 77, 791-798 (2009). Production trials by using inert gas instead of pure gas is first done at 1968 (U.S. Pat. No. 3,364,200) by Johnson and Johnson Company. 8-13% oxidation in 16 hours, 18-22% in 72 hours was got to use 20% NO2 gas in CCl4, Freon-113, Freon-11 that is written U.S. Pat. No. 6,627,749 B1, WO 2007/085364.
J&J Co. have still supplied to the world, to produce oxidised cellulose by NO2 oxidation which is validated by oxidation process. REOXC (Surgicel®) as absorbable hemostatic is marketed. Both clinically and during the operations, it is important hemostatic of external wounding.
Although these important properties, REOXC has disadvantage. Either partly anti-microbiological or biocompatibility or biodegradability is problematic. However it is written on “X. Zhou, X. Huan, US 2010/0298264A1” that degradation time is 5-7 days, absorption time is 8-15 days, —COOH yield is 8-18%, hemostatic time is 2 min; this statement is related with —COOH groups carried the product. In our study if the yield of —COOH is 8-12%, increasing at absorbability and degradability is determined. —COOH yield is 22-24% cause sensitivity and irritability to nervous system, it is disadvantage and undesired status. “Watt et al. EP 1325754A1”. The best hemostatic property —COOH yield must be limited “Stilwell et. al U.S. Pat. No. 5,484,913”. To increasing absorbability and degradability, Ca+2 and Ca-salts can be added to free —COOH.
In this invention Ca-ReOxicell (REOXI-C, Sample3) powder which pH is 4.0-4.5 is prepared and both hemostatic effect and biodegradable features is getting better. To produce powder form is easy but band and cotton-spanch producing is hard to provide this specs. (saferstein, L., et. al. (1992) U.S. Pat. Nos. 5,134,229; 5,134,229; 5,484,913).
In this study and invention, REOXC is prepared with Ca-asetate, Na-asetate and Ca, Na salts. We determined via animal tests that hemostatic properties are decreasing while Na amount and solubility in water is increasing.
1.3. Literatures:    1. Alpaslan, C., Alpaslan, G. H., & Oygur, T. (1997). British Journal of Oral and Maxillofacial Surgery, 35, 129-132.    2. Ashworth, D. R., & Whear, N. M. (2003). British Journal of Oral and Maxillofacial Surgery, 41, 353-354.    3. Breech, L. L., & Laufer, M. R. (2000). Surgicel®. Journal of Pediatric and Adolescent Gynecology, 13, 21-22.    4. Domb, A. J., Kost, J., & Wiseman, D. M. (1998). Handbook of biodegradable polymers. In R. L. Stilwell, M. G. Marks, L. Saferstein, & D. M. Wiseman (Eds.), Oxidized cellulose: Chemistry, processing and medical application (pp. 291-306). BocaRaton: CRC.    5. Loescher, A. R., & Robinson, P. P. (1998). British Journal of Oral and Maxillofacial Surgery, 36, 327-332.    6. Sharma, J. B., & Malhotra, M. (2006 (Surgicel Nu Knit): A case report. Archives of Gynecology, 274, 115-116.    7. Saito, T., Okita, Y., Nge, T. T., Sugiyama, J., & Isogai, A. (2006) Carbohydrate Polymers, 65, 435-440.    8. Stilwell, R. L., Whitmore, E. J., & Saferstein, L. G. (1996). Calcium-modified oxidizedcellulose hemostat. U.S. Pat. No. 5,484,913.    9. Watt, P. W., Harvey, W., & Wiseman, D. (2003). Wound dressing materials comprising collagen and oxidized cellulose. EP 1325754A1.    10. Yackel, E. C., & Kenyon, W. O. (1941). The oxidation of cellulose by nitrogen dioxide., Journal of the American Chemical Society, 64, 121-127.    11. Yin, X., Koschella, A., & Heinze, T. (2009). Regioselectively oxidized 3-O-alkyl ethers of cellulose: Synthesis and characterization. Reactive & Functional Polymers, 69, 341-346.    12. Zhu, L. H., Kumar, V., & Banker, G. S. (2001). Examination of oxidized celluloseas a macromolecular prodrug carrier: Preparation and characterization of an oxidized cellulose-phenylpropanolamine conjugate. International Journal of Pharmaceutics, 223, 35-47.    13. Zimnitsky, D. S., Yurkshtovich, T. L., & Bychkovsky, P. M. (2004). Synthesis and characterization of oxidized cellulose. Journal of Polymer Science Part A: PolymerChemistry, 42, 4785-4791.    14. Zimnitsky, D. S., Yurkshtovich, T. L., & Bychkovsky, P. M. (2006). Adsorption of zwitterionic drugs on oxidized cellulose from aqueous solutions. Reactive & Functional Polymers, 66, 519-525.    15. 8. Harvey, W., Leeuwen, P. V., Hyland, T., & Aitken, W. (2001). Freeze-dried composite materials and processes for the production thereof. U.S. Pat. No. 6,309,454B1.